鉴别外周血与月经血的六重mRNA荧光复合检测体系构建

Construction of a six-plex mRNA fluorescence multiplex identification system for identifying peripheral blood and menstrual blood

目的基于外周血、月经血中特异的mRNA分子标记,建立一种能有效鉴别外周血与月经血的六重mRNA荧光复合检测体系。方法通过文献及数据调研,筛选出在外周血和月经血中特异性表达的mRNA标记分子,设计并验证引物。经过总RNA提取、逆转录、多重PCR(MPCR)扩增、遗传分析仪电泳检测分析等步骤,构建一个基于毛细管电泳技术的六重mRNA荧光复合检测体系,并使用38份样本对体系进行检测验证。结果在外周血样本中检测到外周血标记(HBB、HBA)和管家基因(GAPDH、ACTB)的表达;在月经血样本中检测到所有标记(HBB、HBA、MMP7、MMP10、GAPDH、ACTB)的表达;在非血样本中只检测到管家基因(GAPDH、ACTB)的表达。结论本研究构建的六重mRNA荧光复合检测系统可以有效鉴别外周血与月经血样本。

Objective To establish a six-plex mRNA fluorescent complex system for identifying peripheral blood and menstrual blood based on cell-specific mRNA marker molecules of peripheral blood and menstrual blood. Methods To screen out mRNA markers that are specifically expressed in peripheral blood and menstrual blood through literature and date research, design and validate primers. Constructing a six-plex mRNA-profiling system for the forensic identification of peripheral blood and menstrual blood by extraction of total RNA from the blood, reverse transcription, multiplex PCR(MPCR), genetic analysis, electrophoresis detection and other steps. Then 58 samples are used to test and verify the system. Results Peripheral blood markers(HBB, HBA) and housekeeping genes(GAPDH, ACTB) were detected in peripheral blood samples;All markers(HBB, HBA, MMP7, MMP10, GAPDH, ACTB) were detected in menstrual blood samples;Only the housekeeping genes(GAPDH, ACTB) were detected in non-blood samples. Conclusion The six-plex mRNA fluorescent complex identification system can effectively identify peripheral blood and menstrual blood.